Department of Microbiology and Cell Biology, Montana State University, Bozeman, Montana, USA.
Department of Molecular Cell Biology, University of California Merced, Merced, California, USA.
Immunol Invest. 2024 Oct;53(7):1001-1012. doi: 10.1080/08820139.2024.2382805. Epub 2024 Aug 8.
The field of immunology has undoubtedly benefited from the use of cell lines for immunological studies; however, due to the "immortal" nature of many cell lines, they are not always the best model. Thus, direct collection and culture of primary cells from model organisms is a solution that many researchers utilize. To the best of our knowledge, there is not a singular protocol which encompasses the entire process of bone marrow cell collection through cryopreservation and resuscitation of cells from a murine model.
Bone marrow cells were collected from mice with a C57BL6 genetic background. Cells were differentiated using L929 conditioned media. Cells were assessed using a combination of microscopy, differential staining, immunocytochemistry, and trypan blue. Results: Primary murine BMDMs that underwent cryopreservation followed by resuscitation retained a high degree of viability. Furthermore, these BMDMs retained on overall ability to clear .
Primary murine BMDMs that underwent cryopreservation followed by resuscitation retained a high degree of viability. Furthermore, these BMDMs retained on overall ability to clear .
Crypopreserved and resuscitated primary murine BMDMs were viable and retained their pverall clearance ability.
免疫学领域无疑受益于细胞系在免疫学研究中的应用；然而，由于许多细胞系的“永生”性质，它们并不总是最佳模型。因此，许多研究人员直接从模式生物中收集和培养原代细胞。据我们所知，目前还没有一个单一的方案涵盖了从 C57BL6 遗传背景的小鼠中收集骨髓细胞，通过冷冻保存和复苏细胞，从鼠模型中收集骨髓细胞的整个过程。
从具有 C57BL6 遗传背景的小鼠中收集骨髓细胞。使用 L929 条件培养基对细胞进行分化。使用显微镜、差异染色、免疫细胞化学和台盼蓝评估细胞。结果：经过冷冻保存和复苏的原代鼠骨髓巨噬细胞（BMDMs）保持了高度的活力。此外，这些 BMDMs 总体上仍具有清除能力。
冷冻保存和复苏的原代鼠 BMDMs 具有活力，并保留了其总体清除能力。
